Principles of Formation of Collection Funds of Microorganism Strains

Грачева, И. В.; Осин, А. В.; Кутырев, Владимир

doi:10.21055/0370-1069-2021-2-16-23

Cited by 3 publications

(1 citation statement)

References 12 publications

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“…Не исключена опасность возникновения новых случаев сапа в связи с неблагополучием по данному заболе-микроорганизмов в нативном виде не соответствует современным требованиям, так как в процессе многократных пересевов накапливаются мутации, которые приводят к изменению их изначальных биологических свойств [15,16,17,18]. Ввиду того что одним из направлений коллекционной деятельности является оптимизация существующих методов и разработка новых технологий консервации штаммов микроорганизмов, актуально проведение работ по усовершенствованию ранее разработанных методик сохранения аутентичности штаммов [19,20,21,22].…”

Section: Introductionunclassified

Improved production strain maintenance technique for Burkholderia mallei 5584 (Master seed) used for glander diagnostic agent production

Artemeva,

Melnikova,

Zaynullin

et al. 2024

Veterinariâ segodnâ

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One of the aspects important for strain collection maintenance is the optimization of existing methods and development of new techniques for microbial strain preservation, that is why the improvement of previously developed methods for authentic strain preservation is an urgent task. The article provides information on the maintenance of Burkholderia mallei 5584 (Master seed) using previously developed technique, which was supplemented with new stages in accordance with modern requirements for strain collections of highly dangerous disease agents. The previous strain maintenance technique involved its storage in its native state, which facilitated accumulation of genetic mutations and, ultimately modification of bacterial cell properties. To extend the storage time of this strain and to ensure the stability of its biological properties, the freeze-drying method was used. Skimmed milk was used as a cryoprotectant. Freeze-drying was performed under selected conditions. This technique allows for the strain sub-culturing on sensitive models once every 5 years, which is more expedient and safe from an economic and biological point of view. For safe handling of Burkholderia mallei 5584 production strain, an inactivation technique using gamma rays at 30 kGy was developed, which allowed to achieve microbial suspension sterility and preserve the bacterial cell structure. When comparing the previously developed and supplemented techniques, it was found that the improved technique of Burkholderia mallei 5584 (Master seed) maintenance makes it possible to avoid the loss of its biological properties needed for the production of high-quality laboratory diagnostic agents used for timely disease detection in susceptible animals by diagnostic tests.

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Section: Introductionunclassified

Improved production strain maintenance technique for Burkholderia mallei 5584 (Master seed) used for glander diagnostic agent production

Artemeva,

Melnikova,

Zaynullin

et al. 2024

Veterinariâ segodnâ

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Viability assessment of <i>Salmonella enterica</i> subsp. <i>enterica</i> serovar Abony NCTC 6017 in ampoules and vials by the accelerated shelf-life testing method

Voropaev,

Cidipova,

Fadeikina

et al. 2024

Biological Products. Prevention, Diagnosis, Treatment

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INTRODUCTION. A key priority in maintaining a collection of microorganisms is to ensure the stability of characteristics and the viability of microbial cultures during their storage and transport. In addition, some applications of collection strains as control samples require accurate data on the number of viable microbial cells in each sample. Therefore, it is necessary to develop and implement an analytical procedure for predicting the guaranteed shelf life of test strains.AIM. This study aimed to predict the guaranteed shelf life for test strains in a variety of primary packaging by assessing changes in microbial viability under accelerated storage conditions, with the Salmonella enterica subsp. enterica serovar Abony NCTC 6017 strain as a model organism.MATERIALS AND METHODS. The study used lyophilised samples of the S. enterica subsp. enterica serovar Abony NCTC 6017 strain deposited in the National Collection of Pathogenic Microorganisms (NCPM) at the Scientific Centre for Expert Evaluation of Medicinal Products of the Ministry of Health of the Russian Federation. The studied primary packaging types included vacuum-filled borosilicate glass ampoules and 2R lyophilisation vials. The samples were tested for a number of quality attributes (loss on drying, viable cell count, cell viability, colony morphology, and biochemical identification) and subjected to accelerated shelf-life testing at elevated temperatures (35–65 °С).RESULTS. The study did not show any significant differences in the quality of lyophilised samples depending on the type of primary packaging. The authors experimentally determined rate constants for the loss of viability in microbial cultures during storage at elevated temperatures and calculated the rate constants for the storage and transport temperatures and for different types of primary packaging. The predicted time to viable cell count reduction to 10% of the initial level was 19 years for vials and 25 years for ampoules, and the predicted time to 50% viability was 5.8 years for vials and 7.6 years for ampoules.CONCLUSIONS. The results of this study confirm the applicability of different primary packaging (ampoules and vials) for the lyophilisation and storage of microbial test strains. The data obtained can guide further research and contribute to the development of recommendations for the storage of lyophilised strains in various types of packaging.

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Storage of Bacillus anthracis strain using low temperature preservation method

Rodionov,

Ivanova,

Sayfullin

et al. 2024

jour

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Storage of industrially valuable strains of microorganisms is one of the strategic tasks of modern science and the biological industry. The production and timely implementation of therapeutic, preventive and diagnostic drugs against anthrax contributes to maintaining a stable epizootic situation in Russia for this disease. Therefore, the development and study of methods for storing industrially valuable strains of Bacillus anthracis is an important research work that allows preserving and preserving the unique biological properties of the cells of this pathogen. The purpose of this work was to study the preservation of the viability and biological properties of the Bacillus anthracis strain after 12 months of low-temperature preservation. The strain K-STI-79 of the causative agent of anthrax was used in the work. The strain was stored for 12 months at a temperature of minus 70 C in two cryoprotective media: 15% glycerin solution with 15% glucose solution and 30% neutral glycerin solution in saline solution. After 6 and 12 months of storage, the cells were removed from preservation, thawed at 37 C, tenfold dilutions were made in sterile saline solution and seeded to determine the concentration of viable cells. The safety of the biological properties of the pathogen was studied according to MUC 4.2.2413-08. As a result of the work, it was found that the selected cryoprotectors and the temperature regime of storage allowed to keep a high number of cells in a viable state, which did not significantly differ statistically from the initial concentration. The study of the biological properties of the strain showed their compliance with passport data. The conducted work demonstrates the possibility of effective use of the applied scheme of low-temperature preservation of Bacillus anthracis strains for 12 months.

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Principles of Formation of Collection Funds of Microorganism Strains

Cited by 3 publications

References 12 publications

Improved production strain maintenance technique for Burkholderia mallei 5584 (Master seed) used for glander diagnostic agent production

Improved production strain maintenance technique for Burkholderia mallei 5584 (Master seed) used for glander diagnostic agent production

Viability assessment of <i>Salmonella enterica</i> subsp. <i>enterica</i> serovar Abony NCTC 6017 in ampoules and vials by the accelerated shelf-life testing method

Storage of Bacillus anthracis strain using low temperature preservation method

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